Functional Influenza NS1-Cellular Protein Interactions

描述：(改编自研究人员摘要)：这项拟议的研究调查了病毒感染期间甲型和乙型流感NS1蛋白与特定细胞蛋白之间的功能相互作用。NS1蛋白是一种非结构蛋白，对A型病毒来说，它调节转录后的事件。NS Lb蛋白的功能(S)仍不清楚。反向遗传学的最新进展使甲型和乙型流感病毒完全可以从克隆的cDNA中产生。这是产生变异流感病毒和研究流感感染过程中发生的蛋白质相互作用的有力工具。这项拟议的研究利用这项技术以及GST下拉、亲和层析以及突变和缺失分析来识别与NS 1蛋白相互作用的特定细胞蛋白。这些蛋白质的相互作用将为开发更有效的流感抗病毒疗法提供洞察力。具体目标是：1)确定特定细胞蛋白和甲型流感病毒NS 1A蛋白之间的功能相互作用，重点是确定这种相互作用与SRp54的功能。将确定蛋白质结合部位，并分析这种特定蛋白质相互作用的功能。2)确定特定细胞蛋白与B型流感病毒NS lb蛋白之间的功能相互作用。将确定蛋白质结合部位，并分析这些特定蛋白质相互作用的功能。重点将放在NS lb和ISG15之间的相互作用上。

DESCRIPTION: (Adapted from the Investigators abstract): The proposed research investigates functional interactions between the influenza A and B NS1 proteins with specific cellular proteins during viral infection. The NS 1 protein is a non-structural protein that for type A viruses regulates events post-transcription. The function(s) of the NS lB protein remains unclear. Recent advances in reverse genetics allow influenza A and B viruses to be generated entirely from cloned cDNAs. This is a powerful tool for the generation of mutant influenza viruses and the investigation of protein interactions that occur during the course of influenza infection. The proposed research utilizes this technique as well as GST pull-downs, affinity chromatography, and mutation and deletion analysis to identify specific cellular proteins that interact with the NS 1 protein. These protein interactions will provide insight that could allow for the development of more effective influenza antiviral therapies. Specific aims are: 1) Identify functional interactions between specific cellular proteins and the NS 1A protein of influenza A virus, with emphasis on identifying the function of this interaction with SRp54. The protein-binding site will be identified and the function of this specific protein interaction analyzed. 2) Identify functional interactions between specific cellular proteins and the NS lB protein of influenza B virus. The protein-binding sites will be identified and the function of these specific protein interactions analyzed. Emphasis will be on the interaction between NS lB and ISG15.